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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
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- Product number
- 711729 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- LOXL2 Recombinant Polyclonal Antibody (7HCLC)
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 7HCLC
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of LOXL2 was achieved by transfecting U87MG cells with LOXL2 specific validated siRNA (Silencer® select Product # S8261). Western blot analysis (Fig a) was performed using Modified Whole cell extract (1% SDS) from the LOXL2 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-LOXL2 Recombinant Rabbit Polyclonal Antibody (Product # 711729, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Densitometric analysis of this Western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to LOXL2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with cobalt chloride (100 µM for 16 hrs) (Lane 2), U-87 MG (Lane 3), U-87 MG treated with cobalt chloride (100 µM for 16 hrs) (Lane 4), MDA-MB-231 (Lane 5), MDA-MB-231 treated with cobalt chloride (100 µM for 16 hrs) (Lane 6), A549 (Lane 7) and A549 treated with cobalt chloride (100 µM for 16 hrs) (Lane 8). The blots were probed with Anti-LOXL2 Recombinant Rabbit Polyclonal Antibody (Product # 711729, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A 87 kDa band corresponding to LOXL2 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, HeLa cells were fixed and permeabilized for detection of endogenous LOXL2 using Anti-LOXL2 Recombinant Rabbit Polyclonal Antibody (Product # 711729, 5 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) is stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938) and cytoskeletal F-actin (red) staining using Rhodamine Phalloidin (Product # R415, 1:300) Panel a-d) shows representative un-treated cells that were stained for detection and localization of LOXL2 protein (green) with less signal. Panel e-h) clearly demonstrate enhanced perinuclear localisation of LOXL2 in cells treated with Cobalt chloride (100µM 16h). The images were captured at 60X magnification.
